Team:HKUST/future work



Future Work for iGEM 2010 HKUST team

  1. Transform Lactobacillus plantarum WCFS1 with our existing constructs [agrC-mCherry/agrC-plnB-mCherry in pMG36e]. Use fluorescence microscope to examine whether the chimeric AIP receptor localizes on Lactobacillus cell membrane.

  2. We will continue our "chimeric AIP receptor functionality testing" experiments in two Lactobacillus species, L. plantarum WCFS1 and L. sakei Lb790. We have already built the part [agrC/agrC-plnB in pMG36e] of our original design and we hope to complete the rest [plnA promoter – gusA report unit] soon. After obtaining the final construct [plnA promoter – gusA reporter – P32 promoter – agrC/agrC-plnB], we will transform the pMG36e shuttle vector into respective Lactobacillus cells. Finally, we will apply gusA reporter assay to determine whether the fusion AIP receptor functions as expected.

  3. Migrate the existing constructs "RIP synthesis and secretion cassette" [SP+DD13-RIP] from pBluescript to shuttle vector pMG36e.

  4. Perform functionality test to examine RIP secretion efficiency in L. plantarum.

  5. Conduct bio-assay of the reporter plasmid and estimate the inhibition efficiency of DD13-RIP against S. aureus growth.

  6. Combine constructs from two modules – "AIP Sensor on Lactobacillus Cell Membrane" and "RIP Synthesis and Secretion in Lactobacillus" – together. We will then transform the combined construct [plnA promoter – RIP synthesis and secretion cassette – P32 promoter – agrC/agrC-plnB] into L. plantarum and examine if our initial design works as expected.